Phase-II Drug Metabolism Pharmaceutical Medicinal Chemistry-I Dr. Bilal Al-Jaidi Assistant Professor in Medicinal Chemistry and Drug Design Faculty of Pharmacy, Philadelphia University-Jordan Email: bjaidi@philadelphia.edu.jo

Learning Outcomes At the end of this lesson, students will be able to: Define what Phase-II drug metabolism mean. Demonstrate the different routes of Phase-II metabolism in body. Explain the effect of Drug Metabolism on drug’s duration of action and activity. Classify mechanisms of drug metabolism-Phase-II and enzymes involved. Outline the role of drug metabolism in drug toxicity and carcinogenicity.

Phase-II Drug Metabolism Reactions which conjugate the drug or its phase-I metabolite with a hydrophilic, endogenous species (conjugation reactions). These endogenous compounds are: Glucuronic acid Sulfate group Amino acid (Glycine) Methyl group (as SAM) Acetyl group (as acetyl CoA) Glutathione (tripeptide)

Glutathione Methyl source Sulfate Glucuronic acid Acetyl CoA Glycine

Phase-II Drug Metabolism Involves the following conjugation reactions that are catalyzed by transferase enzymes: Glucuronidation. Sulfation. Amino acid conjugation. Methylation. Acetylation.

Phase-II Drug Metabolism The drug must have a group capable of forming a bond with the endogenous compound. If the drug does not have such group, it will undergo phase-I metabolism, then phase-I metabolite will be conjugated during phase-II reactions.

EXCRETION PHASE-II PHASE-I DRUG

Phase-II Drug Metabolism Most of the time gives metabolite that is: More polar Non-toxic Pharmacologically inactive

X = O, N, S Phenol, alcohol and carboxylic acid Amine (aliphatic and aromatic) Thiol

Sometimes: Phase-II gives: Less polar conjugate: Toxic metabolite: Methylated drug Acetylated drug Toxic metabolite: Some sulfate conjugates Some Acetylated metabolites.

Glucuronidation Involves conjugation of drug with glucuronic acid. IT must be activated to be a good leaving group Hydroxyl group is a bad leaving group

Mechanism of conjugation

Examples Estradiol

Examples Morphine is extensively metabolized into glucuronide conjugate

Example of C-glucuronidation

Example of C-glucuronidation Nucleophilic attack Because this H-atom is acidic

Possible groups for glucuronide conjugation

Possible groups for glucuronide conjugation

Sulfation (sulfate conjugation) Occurs primarily for phenols and occasionally for alcohols, arylamines, and N-hydroxy compounds catalyzed by sulfotransferase enzyme. Sulfotransferase is available mainly in liver, but can be found in kidney, intestine and other tissues

Sulfation Also occur for endogenous compounds, such as steroids, thyroxin, catecholamine and heparin.

Sulfation The first step is the bioactivation of inorganic sulfate by enzyme called ATP sulforylase to give the coenzyme 3'-phosphoadosine-5’-phosphosulfate (PAPS)

Sulfation The second step here is the transfer of sulfate group from the coenzyme PABS to the acceptor drug by nucleophilic attack:

As in all conjugation reactions: The endogenous polar group must be activated and converted into electrophilic derivative. Then the drug nucleophilic group will attack the reactive form get the polar, ionizable endogenous molecule.

Examples of sulfation

Most of the time, sulfate conjugation will give non toxic, polar and easily excreted metabolite. Sometimes, the sulfate conjugate will be converted into toxic metabolite when the sulfate leaves the compound leaving a highly electrophilic intermediate.

Hepato and Nephrotoxicity

Acetylation Is a reaction of amino groups involving the transfer of acetyl group to: An aromatic or aliphatic primary amine Amino acids Hydrazine Hydrazide Sulfonamides Secondary and tertiary amines are not acetylated.

Acetylation The acetylated drug is generally inactive and noon toxic. In contrast to other metabolic transformations, acetylation ends with less polar metabolite compared to the parent drug In some cases, the acetylated metabolite will be as active as the original drug.

Example of active acetylated metabolite

Example of toxic acetylated metabolite Liver damage Highly reactive specie

Example of toxic acetylated metabolite Cause crystalluria….kidney failure Precipitate out in the urine Less water soluble than sulfathiazole

Mechanism of acetylation Acetyl CoA is the activated carrier for acetyl group The reaction catalyzed by the soluble acetyltransferase enzyme: Mainly found in liver Might found in lung, spleen, GIT and red blood cells

Mechanism of acetylation

Examples of acetylation

Acetylation The rate of acetylation is mainly affected by the existence of genetic polymorphism. Two acetylator phenotypes: Slow acetylators: tend to accumulate higher blood concentrations of un-acetylated drugs...toxicity (isoniazid… peripheral nerve damage and liver damage) Fast acetylators: eliminate drugs more rapidly, at the same time can form toxic metabolite very fast. Egyptians and western Europeans are slow acetylator Eskimos and Asians are rapid acetylator

An introduction to Medicinal Chemistry by Graham L. Patrick An introduction to Medicinal Chemistry by Graham L. Patrick. 4th edition, Oxford, 2009 Wilson and Gisvolds text book of organic medicinal and pharmaceutical chemistry by John H. Black and John M. Beale, jr. 12th edition, Lippincott Williams and Wilkings 2011. Foyes principle of medicinal chemistry by David H. Williams, Thomas L. Leuke, Williams O. Foye. Lippincott William and Wilkins. 7th edition, 2013.

The End